phospho igf 1r elisa kit  (R&D Systems)

Journal: Biomedicines

Article Title: Regulation of Soluble E-Cadherin Signaling in Non-Small-Cell Lung Cancer Cells by Nicotine, BDNF, and β-Adrenergic Receptor Ligands

doi: 10.3390/biomedicines11092555

Figure Lengend Snippet: Immunodepletion of sE-cad inhibited activation of EGFR and IGF-1R in A549 cells. Cells were incubated in serum-free media for 12 h, then treated for 72 h with 300 μL of media (0.5 μg/μL) immunodepleted using either hIgG (ID hIgG) or sE-cad (ID sE-cad) (Methods) prepared from cells either untreated (Control) or treated with BDNF (5 nM), nicotine (Nic, 1 µM), or epinephrine (Epi, 100 nM), as described in the legend. The phospho/total EGFR assay ( A , B ) and the phospho/total IGF-1R assay ( C , D ) were then carried out (Methods). Data were expressed as fold change relative to ID hIgG control untreated cells using the GraphPad 9.5.1 software (n = 5). Asterisks indicate a statistically significant difference between each treatment ID from sE-cad relative to the same treatment ID using hIgG, ** p < 0.0l. Please note the differences in scale. Asterisks indicate a statistically significant difference from the corresponding control for each cell line, using the Mann–Whitney test. For comparing three or more groups, the Kruskal–Wallis test, followed by Dunn’s multiple comparison test, was performed. The absence of asterisks indicates no significance; ** p < 0.0l.

Article Snippet: The Phospho-IGF-1R (Tyr1165/1166) and Total IGF-1R ELISA kits (RayBiotech, Peachtree Corners, GA, USA) were used to quantitate activated (phosphorylated) IGF-1R according to the manufacturer’s instructions and as previously reported [ ].

Techniques: Activation Assay, Incubation, Software, MANN-WHITNEY, Comparison

Journal: Biomedicines

Article Title: Regulation of Soluble E-Cadherin Signaling in Non-Small-Cell Lung Cancer Cells by Nicotine, BDNF, and β-Adrenergic Receptor Ligands

doi: 10.3390/biomedicines11092555

Figure Lengend Snippet: Summary of the findings from this study using A549 cells. The levels of MMP9 increased upon cell treatment with nicotine, epinephrine (Epi), or BDNF. Increased MMP9 levels lead to an increase in the levels of sE-cad in the media, increasing activation of EGFR and IGF-1R, enhancing PI3K and ERK1/2 signaling, and inhibiting p53 activity, resulting in decreased apoptosis and increased cell survival.

Article Snippet: The Phospho-IGF-1R (Tyr1165/1166) and Total IGF-1R ELISA kits (RayBiotech, Peachtree Corners, GA, USA) were used to quantitate activated (phosphorylated) IGF-1R according to the manufacturer’s instructions and as previously reported [ ].

Techniques: Activation Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Cisplatin Resistance in Lung Cancer Cells by Nicotine, BDNF, and a β-Adrenergic Receptor Blocker

doi: 10.3390/ijms232112829

Figure Lengend Snippet: Phospho/Total IGF-1R ratio increased by cell treatment with epinephrine or nicotine and decreased by cell treatment with propranolol, while blocking IGF-1R activation by PPP increased cell sensitivity to cisplatin. ( A ) Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h. The following day, the cell monolayers were incubated in serum-free media for 24 h (Control), then incubated without or with FBS for 72 h. The phospho/total IGF-1R assay ( A ) was carried out on the same amount of protein (25 µL of 400 µg/mL total protein) of the cell lysate as described in the Methods section. The phospho/total IGF-1R assay was also carried out on A549 and H1299 cells grown as above then incubated in serum-free media for 72 h in the absence or presence of epinephrine (Epi, 100 nM), propranolol (Prop, 1 µM), nicotine (Nic, 1 µM), BDNF (5 nM), PPP (5 µM) or in combination without ( B , C ) or with cisplatin ( D , E ). Cell viability ( F ) and apoptosis ( G ) were determined in the presence of PPP (5 µM) without or with 10 µM cisplatin when using A549 cells and 30 µM cisplatin when using H1299 cells, as described in the Methods section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to untreated control cells (Control) using the GraphPad 9.4.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks (*) indicate a statistically significant difference from the corresponding control for each cell line, Mann–Whitney test, while the absence of asterisks indicates no significance. * p < 0.05, ** p < 0.01.

Article Snippet: Activated (phosphorylated) IGF-1R was quantitated using the Phospho-IGF-1R (Tyr1165/1166) and Total IGF-1R ELISA kit (RayBiotech, Norcross, GA, USA), according to the manufacturer’s instructions.

Techniques: Blocking Assay, Activation Assay, Incubation, Software, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Enhanced anti-metastatic bioactivity of an IGF-TRAP re-engineered to improve physicochemical properties

doi: 10.1038/s41598-018-35407-2

Figure Lengend Snippet: Pharmacokinetic analysis reveals a more rapid clearance of IGF-TRAP 3.1 from the circulation following bolus i.v. injections in three mouse strains. C57Bl/6, Balb/c and nu/nu female mice were tail-vein injected with 10 mg/kg of the indicated IGF-TRAPs. Blood was collected at the indicated time intervals for measurement of circulating IGF-TRAP proteins in the serum using soluble IGF-1R ELISA assay. Data are based on 3 mice bled at the same time intervals. Shown in ( A – C ) are mean serum concentrations values (±SEM) for C57Bl/6, Balb/c and nu/nu mice respectively. Shown in ( D ) are calculated clearance and area under the serum concentration-time curve values determined using the Phoenix Win-Nonlin Software v6.2. a T last – time of the last measured plasma concentration. b AUC 0-last – area under the concentration-time curve up to the last measured plasma concentration. c CL – total body clearance.

Article Snippet: Following a 15 min incubation at 37 °C to activate the IGF-1R, the cells are lysed (in HEPES lysis buffer) and the lysate transferred to a microtiter plate pre-coated with an IGF-1R capture antibody (Human phospho-IGF-1R ELISA kit, R&D Systems, Minneapolis, MN) and probed with an HRP-conjugated anti-phospho-tyrosine antibody.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Software, Clinical Proteomics